Role of MT-MMPs in Renal Development

MT-MMP 在肾脏发育中的作用

• 批准号: 7489677
• 负责人: ROY ZENT
• 金额: $ 2.63万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-03-01 至 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7489677
• 关键词: Academic Medical Centers; Adhesions; Adult; Basement membrane; Binding; Cell physiology; Cells; Cities; Class; Cleaved cell; Data; Defect; Deformity; Development; Duct (organ) structure; Embryo; Endopeptidases; Enzymes; Extracellular Matrix; Extracellular Matrix Degradation; Extracellular Matrix Proteins; Face; Family; GPI Membrane Anchors; Gel; Human Resources; In Vitro; Instruction; Kidney; Knockout Mice; Last Name; Matrix Metalloproteinases; Membrane; Metalloproteases; Metanephric Diverticulum; Morphogenesis; Mus; Names; Nephrons; Numbers; Peptide Hydrolases; Phenotype; Physiological; Play; Principal Investigator; Printing; Proteins; Research Personnel; Research Project Grants; Role; Sequence Homology; Site; Structure; System; Testing; Universities; base; collecting tubule structure; extracellular; human MMP14 protein; in vivo; laminin-5; member; migration; mutant; programs; reconstitution; research study

Matrix metalloproteinases (MMP) are a multigenic family of more than 20 proteolytic enzymes involved in the degradation of extracellular matrix (ECM) components. Most MMPs are secreted as soluble enzymes into the extracellular milieu; however some are membrane-bound and called the membrane-targeted metalloproteinases- MMPs. Despite in vitro evidence that soluble MMPs play an important role in renal development, no significant renal phenotypes have been described in mice lacking this class of MMPs. In contrast, we have found that mice null for MT1-MMP have dysplastic dysgenic kidneys due to decreased cleavage of ECM components, particularly laminin 5 (Ln-5). More recently we observed that MT4-MMP null mice have hypoplastic dysgenic kidneys, due to an undetermined mechanism. Based on these data we hypothesize that MT-MMPs_ namel F MTI- and MT4-MMPs play a critical role in normal renal development. The role of these enzymes in normal renal development will be determined in the following 3 aims. 1) We will determine whether Ln-5 cleavage is required for nomaal kidney devclopmcnt by a) crossing thc Ln-5-null mouse with the MT1-MMP-null mouse and comparing the phenotype of the double null mutants with the single Ln-5-null and MT1-MMP null mice to see if the renal phenotype of the MT1-MMP-null mouse can be rescued; b) isolating whole embryonic kidneys and collecting duct cells from the Ln- 5, MT1-MMP and Ln-5/MT1-MMP null mice and determine their ability to undergo branching movphogenesis. In the tubulogenesis experiments the gels will be reconstituted with different cleavage products of Ln-5 to determine which specific domains of Ln-5 are critical for branching morphogenesis. 2) We will determine how lack of MT4- MMP results in hypoplastic and dysgenic kidneys by a) analyzing embryonic and adult kidneys from wild type and MT4-MMP-null mice and b) isolating collecting duct cells from wild type and MT4-MMP-null mice to determine the effects of lack of this enzyme on ECM-dependent cellular functions such as migration, adhesion and tubulogeneisis. In aim 3 we will determine the relevant ECM substrates for MT1- and MT4-MMP by determining a) in vitro the cleavage products of purified renal basement membrane components by purified MT1- and MT4-MMPs alone or in combination with soluble MMPs and b) in vivo whether there are differences in the composition of the renal basement membranes of the MT 1- and MT4-MMP null mice compared to their wild type counterparts.

基质金属蛋白酶（MMP）是一个多基因家族，该家族由20多种蛋白水解酶，参与细胞外基质（ECM）成分降解。大多数MMP被分泌为可溶性酶 细胞外环境；但是，有些是膜结合的，称为膜靶向金属蛋白酶-MMP。尽管有证据表明可溶性MMP在肾脏发展中起重要作用，但没有显着 在缺乏这类MMP的小鼠中已经描述了肾脏表型。相比之下，我们发现MT1-MMP的无效小鼠由于ECM成分的裂解降低而患有发育不良的肾脏肾脏，特别是 层粘连蛋白5（LN-5）。最近，我们观察到，由于不确定的机制，MT4-MMP无效小鼠患有肾上腺素肾脏肾脏肾上腺素障碍性肾脏不佳。基于这些数据，我们假设MT-MMPS_ NAMEL F MTI-和MT4-MMP 在正常的肾脏发展中起关键作用。这些酶在正常肾脏发育中的作用将在以下3个目标中确定。 1）我们将确定是否需要通过a）与MT1-MMP-NULL小鼠越过THC LN-5无效小鼠，并比较单个LN-5-5-NULL和MT1-MMP NULL小鼠，以查看MT1-MMP Null小鼠的表型，以查看肾脏鼠标MT的MMT MMM，是否需要LN-5裂解。 b）从LN-5，MT1-MMP和LN-5/MT1-MMP无效小鼠中分离整个胚胎肾脏并收集导管细胞，并确定其进行分支分支的移动生成的能力。在肾小管生成实验中，将用LN-5的不同裂解产物重构凝胶，以确定LN-5的哪些特定域对于分支形态发生至关重要。 2）我们将通过a）分析野生型和MT4-MMP-NULL小鼠的胚胎和成年肾脏的缺乏MT4-MMP会导致低塑性和失调肾脏导致肾脏肾脏，以及b）分离从野生型和MT4-MMP-NULL小鼠中分离出收集的导管，以确定ECM依赖性依赖性功能的影响。管状。 In aim 3 we will determine the relevant ECM substrates for MT1- and MT4-MMP by determining a) in vitro the cleavage products of purified renal basement membrane components by purified MT1- and MT4-MMPs alone or in combination with soluble MMPs and b) in vivo whether there are differences in the composition of the renal basement membranes of the MT 1- and MT4-MMP与野生型相比，无效的小鼠。